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All microscope settings were set to collect images below saturation and were kept constant for all images taken in one experiment, as previously described [ 26].
A series range for each section was set by determining an upper and lower threshold using the Z/Y Position for Spatial Image Series setting, and confocal microscope settings were established and maintained by Leica and local technicians for optimal resolution.
Confocal microscope settings were adjusted to produce the optimum signal-to-noise ratio.
For each image the same microscope settings were employed to simplify comparison and to avoid overexposure.
Identical camera and microscope settings were employed to allow valid comparison between images of control and macroH2A1 deficient cell populations.
The same microscope settings were used to analyze YFP-ABF3 and YFP-OST1 cellular localization in Arabidopsis transgenic plants.
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It should be noted that, if the microscope settings are optimized, a very faint level of fluorescence can be detected at each stage of development.
Moreover, even if the confocal microscope settings are modified to allow greater sensitivity to detect cellular doxorubicinol, doxorubicinol was found not to be localized to the nucleus in both MCF-7CC12 and MCF-7DOX2-12 MCF-7DOX2-12 MCF-7DOX2-12
The same microscope and settings were used for all samples, and the slides were read by two independent readers.
However, when microscope gain settings were increased to >850 to allow visualisation of cells expressing very low levels of GFP-BRD1, cells with a homogeneous distribution of tagged protein could be observed.
All microscope and camera settings were identical between groups and images were collected at ×60 magnification.
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